Leukemia research, 2007-04, Vol.31 (4), p.515-521
2007
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- Vector-based RNAi approach to isoform-specific downregulation of vascular endothelial growth factor (VEGF)165 expression in human leukemia cells
- Ist Teil von
- Leukemia research, 2007-04, Vol.31 (4), p.515-521
- Ort / Verlag
- England: Elsevier Ltd
- Erscheinungsjahr
- 2007
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Abstract Vascular endothelial growth factor (VEGF) plays a critical role during normal embryonic angiogenesis and also in the pathological angiogenesis that occurs in a number of diseases, including cancer. K562 human leukemia cells overexpress VEGF, with a shift in isoform production from membrane-bound VEGF189 to the more soluble VEGF165. In the present study, three 19 bp reverse repeated motifs targeting exons 5 and 7 boundary of VEGF165 gene sequence with 9 bp spacer were synthesized and cloned into eukaryotic expression plasmid pGenesil-1 containing U6 shRNA promoter and termination signal of RNA polymerase. The recombinant plasmids pGenesil-VR1, pGenesil-VR2, pGenesil-VR3 and pGenesil-con (plasmid containing random DNA fragment) were transfected into K562 cells, respectively, through lipofectamine™ reagent. A vector-based small interfering RNA(SiRNA) inhibited VEGF165 mRNA expression by 72% and protein production by 67% in K562 cells. Human microvascular endothelial cell migration induced by conditioned medium from VEGFsi-transfected K562 cells was significantly less than that induced by conditioned medium from K562 cells and control vector-transfected K562 cells. Furthermore, the VEGF shRNA dramatically suppressed tumor angiogenesis and tumor growth in a K562 s.c. xenograft model. Vessel density as assessed by vWF immunohistochemical analysis was also decreased. This strategy provides a novel tool to study the function of various VEGF isoforms and may contribute to VEGF-specific treatment in cancer.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0145-2126eISSN: 1873-5835DOI: 10.1016/j.leukres.2006.09.011Titel-ID: cdi_proquest_miscellaneous_68994273
- Format
- –
- Schlagworte
- Animals, Apoptosis, Blotting, Western, Cell Movement, Cell Proliferation, Down-Regulation, Gene Expression Regulation, Neoplastic, Gene therapy, Genetic Vectors - genetics, Hematology, Oncology and Palliative Medicine, Humans, Immunoenzyme Techniques, Isoform, K562 Cells, Leukemia cell, Leukemia, Experimental - genetics, Leukemia, Experimental - metabolism, Leukemia, Experimental - prevention & control, Mice, Mice, Nude, Neovascularization, Pathologic - genetics, Neovascularization, Pathologic - metabolism, Neovascularization, Pathologic - pathology, Protein Isoforms, RNA interference, RNA, Messenger - genetics, RNA, Messenger - metabolism, RNA, Small Interfering - genetics, RNA, Small Interfering - metabolism, Survival Rate, Transfection, Vascular Endothelial Growth Factor A - genetics, Vascular Endothelial Growth Factor A - metabolism, VEGF, Xenograft Model Antitumor Assays - methods